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Identification of stemness and differentially expressed genes in human cementum-derived cells

  • Lee, EunHye (Dental Research Institute, Seoul National University) ;
  • Kim, Young-Sung (Department of Periodontics, Asan Medical Center, University of Ulsan College of Medicine) ;
  • Lee, Yong-Moo (Department of Periodontology and Dental Research Institute, Seoul National University School of Dentistry) ;
  • Kim, Won-Kyung (Department of Periodontics, Asan Medical Center, University of Ulsan College of Medicine) ;
  • Lee, Young-Kyoo (Department of Dentistry, Uijeongbu Eulji Medical Center, Eulji University) ;
  • Kim, Su-Hwan (Department of Periodontics, Asan Medical Center, University of Ulsan College of Medicine)
  • 투고 : 2021.04.23
  • 심사 : 2021.07.09
  • 발행 : 2021.10.30

초록

Purpose: Periodontal treatment aims at complete regeneration of the periodontium, and developing strategies for periodontal regeneration requires a deep understanding of the tissues composing the periodontium. In the present study, the stemness characteristics and gene expression profiles of cementum-derived cells (CDCs) were investigated and compared with previously established human stem cells. Candidate marker proteins for CDCs were also explored. Methods: Periodontal ligament stem cells (PDLSCs), pulp stem cells (PULPSCs), and CDCs were isolated and cultured from extracted human mandibular third molars. Human bone marrow stem cells (BMSCs) were used as a positive control. To identify the stemness of CDCs, cell differentiation (osteogenic, adipogenic, and chondrogenic) and surface antigens were evaluated through flow cytometry. The expression of cementum protein 1 (CEMP1) and cementum attachment protein (CAP) was investigated to explore marker proteins for CDCs through reverse-transcription polymerase chain reaction. To compare the gene expression profiles of the 4 cell types, mRNA and miRNA microarray analysis of 10 samples of BMSCs (n=1), PDLSCs (n=3), PULPSCs (n=3), and CDCs (n=3) were performed. Results: The expression of mesenchymal stem cell markers with a concomitant absence of hematopoietic markers was observed in PDLSCs, PULPSCs, CDCs and BMSCs. All 4 cell populations also showed differentiation into osteogenic, adipogenic, and chondrogenic lineages. CEMP1 was strongly expressed in CDCs, while it was weakly detected in the other 3 cell populations. Meanwhile, CAP was not found in any of the 4 cell populations. The mRNA and miRNA microarray analysis showed that 14 mRNA genes and 4 miRNA genes were differentially expressed in CDCs vs. PDLSCs and PULPSCs. Conclusions: Within the limitations of the study, CDCs seem to have stemness and preferentially express CEMP1. Moreover, there were several up- or down-regulated genes in CDCs vs. PDLSCs, PULPSCs, and BMSCs and these genes could be candidate marker proteins of CDCs.

키워드

과제정보

This study was supported by a grant (2013-541) from the Asan Institute for Life Sciences, Seoul, Korea.

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